LOCUS       sequence-424957-       10646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector to clone sgRNA(s) into the CROPseq-multi construct with
            Puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10646)
  AUTHORS   Walton RT, Qin Y, Blainey PC
  TITLE     CROPseq-multi: a versatile solution for multiplexed perturbation and
            decoding in pooled CRISPR screens
  JOURNAL   bioRxiv 2024.03.17.585235
REFERENCE   2  (bases 1 to 10646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.17.585235"
COMMENT     Sequence Label: CROPseq-multi-Puro (pRW1441)
FEATURES             Location/Qualifiers
     source          1..10646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1489..1606
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1745..2923
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1976..2914
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2871..2891
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2936..3535
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2936..2955)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3432..3452
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3551..4139
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3604..3624)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4022..4033)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(6320..6568)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6378..6397)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6548..6568)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     LTR             6615..6795
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6873..6994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6910..6929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6964..6983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7034..7169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7189..7208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7190..7208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7218..7235)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7218..7234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7227..7249)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7376..7831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7463..7482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7673..7694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7857..7961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7962..8030,8031..8822)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8180..8199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8993..9581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9482..9501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9735..9752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9869..9890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9905..9910,9911..9928,9929..9935)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9943..9959
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9948..9970
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9967..9983
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9967..9983
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10002..10022
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10004..10022
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        10050..10429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10431..10629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10584..10604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     1501 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     1561 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     1621 ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa
     1681 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     1741 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     1801 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     1861 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     1921 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     1981 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2041 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     2101 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     2161 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     2221 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     2281 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     2341 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     2401 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     2461 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     2521 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     2581 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     2641 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     2701 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     2761 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     2821 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     2881 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac
     2941 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3001 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3061 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3121 caaggtgtgg gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc cggagagcgt
     3181 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3241 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3301 gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3361 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3421 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3481 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg
     3541 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3601 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3661 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3721 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3781 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3841 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3901 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3961 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4021 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4081 tccgcgtgtt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     4141 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     4201 aaggggggac tggaagggct aattcactcc caacgaagag aagatttcac cattatcgtt
     4261 tcagacccac ctcccaaccc cgaggggacc cagaaaaaaa gcaccgactc ggtgccactt
     4321 tttcaagttg ataacggact agccttattt caacttgcta tgctgttaga gacgtacaaa
     4381 aaagagcaag aagctaaaaa agatttaaaa attattttta gcgcagttaa tggaacagga
     4441 actaaattta ccccaaaaat attacgtgaa tcaggatata acgttattga ggttgaagag
     4501 catgcatttg aagatgaaac atttaaaaat gttgtaaatc caaatccaga atttgatcct
     4561 gcatgaaaaa taccgcttga atatggtatt aaacatgatg cagatattat tattatgaat
     4621 gacccagatg ctgacagatt tggaatggca ataaaacatg atggtcattt tgtaagatta
     4681 gatggaaatc aaacaggacc aattttaatt gattgaaaat tatcaaatct aaaacgctta
     4741 aatagcattc caaaaaatcc ggctctatat tcaagttttg taacaagtga tttgggtgat
     4801 agaatcgctc atgaaaaata tggagttaat attgtaaaaa ctttaactgg atttaaatga
     4861 atgggtagag aaattgctaa agaagaagat aacggattaa attttgtttt tgcttatgaa
     4921 gaaagttatg gatatgtaat tgatgactca gctagagata aagatggaat acaagcttct
     4981 atattaatag cagaggctgc ttgattttat aaaaaacaaa ataaaacatt agtagactat
     5041 ttagaagatt tatttaaaga aatgggtgca tattacactt tcactttaaa cttgaatttt
     5101 aaaccagaag aaaagaaatt aaaaattgaa ccattaatga aatcattgag agcaacaccc
     5161 ttaactcaaa ttgctggact taaagttgtt aatgttgaag actacatcga tggaatgtat
     5221 aatatgccag gacaagactt actaaaattt tatttagaag ataagtcatg atttgctgtt
     5281 cgcccaagtg gaactgaacc taaactaaaa atttatttta taggtgttgg tgaatctgtt
     5341 caaaacgcta aagttaaagt agacgaaatt attaaagaat taaaattaaa aatgaatata
     5401 taggagaaaa aatgaaacta aacaaatata tagatcacac attattaaaa caagatgcta
     5461 cgaaagctga aattaaacaa ttatgtgatg aagcaattga atttgatttt gcaacagttt
     5521 gtgttaattc atattgaaca agctattgta aagaattatt aaaaggcaca aatgtaggaa
     5581 taacaaatgt tgtaggtttt cctctaggtg catgcacaac agctacaaaa gcattcgaag
     5641 tttctgaagc aattaaagat ggtgcaacag aaattgatat ggtattaaat attggtgcat
     5701 taaaagacaa aaattatgaa ttagttttag aagacatgaa agctgtaaaa aaagcagctg
     5761 gatcacatgt tgttaaatgt attatggaaa attgtttatt aacaaaagaa gaaatcatga
     5821 aagcttgtga aatagctgtt gaagctggat tagaatttgt taaaacatca acaggatttt
     5881 caaaatcagg tgcaacattt gaagatgtta aactaatgaa gtcagttgtt aaagacaatg
     5941 ctttagttaa agcagctggt ggagttagaa catttgaaga tgctcaaaaa atgattgaag
     6001 caggagctga ccgcttagga acaagtggtg gagtagctat tattaaaggt gaagaaaaca
     6061 acgcgagtta ctaaaactag cgttttttta ttttgctcat ttttattaaa agtttgcaaa
     6121 aaggaacata aaaattctaa ttattgatac taaagttatt aaaaagaaga ttttggttga
     6181 ttttataaag gtcatagaat ataatatttt agcatgtgta ttttgtgtgc tcatttacaa
     6241 ccgtctctgc attggccggg aatcgaaccc gggcctcccg cgtggcaggc gagaattcta
     6301 ccactgaacc accaatgccg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat
     6361 cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa
     6421 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
     6481 tgtttacagt caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat
     6541 atgaaggaat catgggaaat aggccctcaa gcttggcgta actagatctt gagacactgc
     6601 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6661 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6721 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6781 ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac
     6841 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6961 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     7021 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7081 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     7141 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     7201 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     7261 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     7321 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     7381 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7441 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7501 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7561 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7621 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7681 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7741 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7801 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     7861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     8041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     8101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     8161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     8221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     8281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     8341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     8401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     8461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     8521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     8581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     8641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     8701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     9001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     9061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     9121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     9181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     9241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     9301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     9361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     9421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     9481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     9541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     9601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     9661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     9721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     9781 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     9841 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     9901 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     9961 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
    10021 gggaacaaaa gctggagctg caagcttaag acattgatta ttgactagtt attaatagta
    10081 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
    10141 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
    10201 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
    10261 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
    10321 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
    10381 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
    10441 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
    10501 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
    10561 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
    10621 tataagcagc gcgttttgcc tgtact
//
